ACE UHPLC and HPLC Columns
Decarboxylation of Sirohaem by Sirohaem Decarboxylase
Column: ACE 5 AQ
Dimensions: 150 x 2.1 mm
Part Number: ACE-126-1502
Mobile Phase: A: 0.1% TFA in H2O
B: MeCN
Gradient: Time (mins) %B
0 5
6 20
25 30
35 100
40 100
Flow Rate: 0.2 mL/min
Detection: DAD, 380 nm
Defensins (Human) in Saliva Matrix
64
Sirohaem
Profile of sirohaem decarboxylase after incubation 3
of sirohaem with purified D. sulfuricans AhbA
12,18-Didecarboxysirohaem
100
%
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Application #AN3830
Conditions Analytes
1. Sirohaem
2. Monodecarboxysirohaem
3. 12,18-Didecarboxysirohaem
1
2
3
10 12 14 16 18 20 22 24 26 28 30
Time - Minutes
1
2
Profile of sirohaem decarboxylase after incubation
of sirohaem with purified D. sulfuricans AhbB
Palmer DJ, Schroeder S, Lawrence AD, Deery E, Lobo SA, Saraiva LM, McLean KJ, Munro AW, Ferguson SJ, Pickersgill RW, Brown DG, Warren MJ. The
structure, function and properties of sirohaem decarboxylase – an enzyme with structural homology to a transcription factor family that is part of an alternative
haem biosynthesis pathway. Molecular Microbiology (2014) 93(2), 247-261. doi:10.1111/mmi.12656
Column: ACE UltraCore 2.5 SuperC18
Dimensions: 50 x 3.0 mm
Part Number: CORE-25A-0503U
Mobile Phase: A: 0.1% formic acid in H2O
B: 0.1% formic acid in MeCN
Gradient: Time (mins) %B
0 2
2 2
17 50
19 95
20 95
Flow Rate: 0.6 mL/min
Detection: Synapt G1 QToF +ESI MS
Sampling cone voltage: 40 V
Source temperature: 150 °C
Capillary voltage: 4.8 kV
Extraction cone voltage: 41 kV
Desolvation temperature: 500 °C
Acquisition: 100-2000 m/z
Sample: SPE on C18
Application #AN1270
Conditions Defensin Human Neutrophil Peptides
1. HNP-1
(30 amino acid residues)
2. HNP-2
(29 amino acid residues)
3. HNP-3
(30 amino acid residues)
TIC
Area of
interest
TIC
2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00
0
Extracted ion current chromatograms
(sum of multiply protonated ions M+3H3+, M+4H4+ and M+5H5+)
100
%
0
10.00 12.00
10.00 12.00
10.00 12.00
100
%
0
100
%
0
1
2
3
1
2
3
100
%
0
100
%
0
100
%
0
200 400 600 800 1000
m/z
m/z
m/z
1200
M+5H5+
689.3223
M+4H4+
861.4352
M+3H3+
1147.8999
M+5H5+
674.9163
M+4H4+
843.6677
M+3H3+
1124.5613
M+5H5+
698.1338
M+4H4+
872.4535
M+3H3+
1162.9043
Time - Minutes
Reproduced with permission of School of Science, University of Greenwich, UK
Mass spectra